绵羊消化道线虫病是严重危害养羊业生产的一类寄生虫疾病，目前使用化学驱虫药是防治该病的主要手段。然而，伴随着耐药虫株的不断产生、动物性食品的药物残留和环境污染等问题日益凸现，迫切需要寻求一种新的动物疫病-环境友好防控方法。利用线虫的自然天敌-捕食线虫性真菌及其分泌性胞外蛋白酶来防控动物线虫病是一种有应用前景的生物防控方法。本项目拟以捕食线虫性真菌的代表菌-少孢节丛孢菌为研究对象，利用染色体DNA步移技术扩增该菌丝氨酸蛋白酶Aoz1基因5'上游侧翼序列，通过系列缺失突变和DNA凝胶阻滞技术鉴定该基因启动子及顺式调控元件；利用根癌农杆菌介导的真菌转化技术（ATMT）构建Aoz1基因启动子缺失和双启动子的重组少孢节丛孢菌，分析重组菌Aoz1基因表达水平及捕食线虫活性，验证Aoz1基因启动子功能，揭示少孢节丛孢菌Aoz1基因表达调控的机制，为研发具有自主知识产权的绵羊线虫病生物防控制剂提供科学依据。

Sheep gastrointestinal nematodiasis is one of animal parasitic diseases, which is seriously harmful to the sheep industry. The wide use of chemical anthelmintics is still the main means to prevent and control the disease. However, with the increasing problems such as drug-resistant strains of insects, drug residues in animal foods and environmental pollution, it is urgent to seek a scientific way to deal with the relationship between control of animal diseases and environmental protection. The use of the natural enemies of nematode-trapping fungi and it's secreted extracellular protease is now considered a promising alternative for the biological control of gastrointestinal nematodiasis. Therefore, it necessary to explore molecular mechanisms on expression of nematode-trapping functional genes. On the basis of preliminary studies, representative of the nematode-trapping fungi-Arthrobotrys oligospora was studied in the project. 5' flanking regions of serine protease Aoz1 gene will be amplified by chromosomal DNA walking technique, then promoter and its regulatory sequences of the gene are analyzed and verified by series of deletion mutations and DNA gel retardation for clarifying it's molecular characteristics. The deleted and double promoter of the Aoz1 gene of recombinant Arthrobotrys oligospora were constructed using Agrobacterium tumefaciens mediated transformation (ATMT) technology, respectively, infection activity and genetic stability were analyzed in nematode. The functional of serine protease Aoz1 gene promoter of Arthrobotrys oligospora was further verified. The elucidation of the molecular mechanisms on expression regulation of Aoz1 gene will provide a scientific basis for development of biological agents with independent intellectual property rights to prevent and control sheep gastrointestinal nematodiasis.